Journal: Experimental and Therapeutic Medicine

Article Title: F-box and WD repeat-containing protein 7 ameliorates angiotensin II-induced myocardial hypertrophic injury via the mTOR-mediated autophagy pathway

doi: 10.3892/etm.2022.11391

Figure Lengend Snippet: FBW7 expression is downregulated in Ang II-induced H9C2 cells and its upregulation activates mTOR-mediated cell autophagy. (A) mRNA expression of FBW7 in the control and Ang II (0.5, 1, 5 and 10 µM) groups. (B) Protein expression level of FBW7 in the control and Ang II (0.5, 1, 5 and 10 µM) groups. Overexpression efficiency of FBW7 in the control, Oe-NC and Oe-FBW7 groups was measured using (C) RT-qPCR and (D) western blotting. (E) Protein expression levels of LC3B-II/I, P62 and Beclin-1 related to autophagy in the control, Ang II, Ang II+Oe-NC and Ang II+Oe-FBW7 groups. (F) Protein levels of p-mTOR and mTOR related to mTOR signaling pathway in the control, Ang II, Ang II+Oe-NC and Ang II+Oe-FBW7 groups. * P<0.05, *** P<0.001. FBW7, F-box and WD repeat-containing protein 7; Ang, angiotensin; Oe, overexpression; NC, negative control; LC3B, microtubule-associated proteins 1A/1B light chain 3B; p-, phosphorylated; -, Ang II treatment without transfection.

Article Snippet: The FBW7 overexpression plasmid (Oe-FBW7) and its negative control (Oe-NC) were obtained from Guangzhou RiboBio Co., Ltd. Oe-FBW7 consisted of a pcDNA3.1 vector containing the full-length FBW7 cDNA sequence, with empty pcDNA3.1 as the negative control (Oe-NC).

Techniques: Expressing, Control, Over Expression, Quantitative RT-PCR, Western Blot, Negative Control, Transfection

Journal: Experimental and Therapeutic Medicine

Article Title: F-box and WD repeat-containing protein 7 ameliorates angiotensin II-induced myocardial hypertrophic injury via the mTOR-mediated autophagy pathway

doi: 10.3892/etm.2022.11391

Figure Lengend Snippet: FBW7 inhibits Ang II-induced apoptosis in H9C2 cells via autophagy. (A) Cell viability of H9C2 cells in the control, Ang II, Ang II+Oe-NC, Ang II+Oe-FBW7 and Ang II+Oe-FBW7+3-MA groups. (B and C) Apoptosis of H9C2 cells in the control, Ang II, Ang II+Oe-NC, Ang II+Oe-FBW7 and Ang II+Oe-FBW7+3-MA groups. Scale bar, 50 µm. (D) Protein levels of Bcl-2, Bax and cleaved caspase 3 associated with apoptosis in the control, Ang II, Ang II+Oe-NC, Ang II+Oe-FBW7 and Ang II+Oe-FBW7+3-MA groups. * P<0.05, ** P<0.01, *** P<0.001. FBW7, F-box and WD repeat-containing protein 7; Ang, angiotensin; Oe, overexpression; NC, negative control; 3-MA, 3-methyladenine; -, Ang II treatment without transfection.

Article Snippet: The FBW7 overexpression plasmid (Oe-FBW7) and its negative control (Oe-NC) were obtained from Guangzhou RiboBio Co., Ltd. Oe-FBW7 consisted of a pcDNA3.1 vector containing the full-length FBW7 cDNA sequence, with empty pcDNA3.1 as the negative control (Oe-NC).

Techniques: Control, Over Expression, Negative Control, Transfection

Journal: Experimental and Therapeutic Medicine

Article Title: F-box and WD repeat-containing protein 7 ameliorates angiotensin II-induced myocardial hypertrophic injury via the mTOR-mediated autophagy pathway

doi: 10.3892/etm.2022.11391

Figure Lengend Snippet: FBW7 inhibits Ang II-induced hypertrophy and fibrosis in H9C2 cells through autophagy. (A) Cell length of H9C2 cells in the control, Ang II, Ang II+Oe-NC, Ang II+Oe-FBW7 and Ang II+Oe-FBW7+3-MA groups. Scale bar, 25 µm. (B) Protein levels of hypertrophic markers β-MHC, BNP and ANF in H9C2 cells in the control, Ang II, Ang II+Oe-NC, Ang II+Oe-FBW7 and Ang II+Oe-FBW7+3-MA groups. (C) Protein levels of α-SMA, Fibronectin, Vimentin Collagen I and Collagen III associated fibrosis in H9C2 cells in the control, Ang II, Ang II+Oe-NC, Ang II+Oe-FBW7 and Ang II+Oe-FBW7+3-MA groups. ** P<0.01, *** P<0.001. FBW7, F-box and WD repeat-containing protein 7; Ang, angiotensin; Oe, overexpression; NC, negative control; 3-MA, 3-methyladenine; β-MHC, β-myosin heavy chain; BNP, brain natriuretic peptide; ANF, atrial natriuretic factor; atrial natriuretic factor; SMA, smooth muscle actin; -, Ang II treatment without transfection.

Article Snippet: The FBW7 overexpression plasmid (Oe-FBW7) and its negative control (Oe-NC) were obtained from Guangzhou RiboBio Co., Ltd. Oe-FBW7 consisted of a pcDNA3.1 vector containing the full-length FBW7 cDNA sequence, with empty pcDNA3.1 as the negative control (Oe-NC).

Techniques: Control, Over Expression, Negative Control, Transfection

Journal: Experimental and Therapeutic Medicine

Article Title: F-box and WD repeat-containing protein 7 ameliorates angiotensin II-induced myocardial hypertrophic injury via the mTOR-mediated autophagy pathway

doi: 10.3892/etm.2022.11391

Figure Lengend Snippet: FBW7 suppresses Ang II-induced inflammation and oxidative stress in H9C2 cells via autophagy. Levels of inflammatory cytokines (A) TNF-α and (B) IL-1β in H9C2 cells in the control, Ang II, Ang II+Oe-NC, Ang II+Oe-FBW7 and Ang II+Oe-FBW7+3-MA groups. (C) Protein levels of iNOS and COX-2 associated with inflammation in H9C2 cells in the control, Ang II, Ang II+Oe-NC, Ang II+Oe-FBW7 and Ang II+Oe-FBW7+3-MA groups. Levels of (D) SOD and (E) MDA associated with oxidative stress in H9C2 cells in the control, Ang II, Ang II+Oe-NC, Ang II+Oe-FBW7 and Ang II+Oe-FBW7+3-MA groups. * P<0.05, ** P<0.01, *** P<0.001. FBW7, F-box and WD repeat-containing protein 7; IL, interleukin; Ang, angiotensin; Oe, overexpression; NC, negative control; 3-MA, 3-methyladenine; iNOS, inducible nitric oxide synthase; COX-2, cyclooxygenase-2; SOD, superoxide dismutase; MDA, malondialdehyde; -, Ang II treatment without transfection.

Article Snippet: The FBW7 overexpression plasmid (Oe-FBW7) and its negative control (Oe-NC) were obtained from Guangzhou RiboBio Co., Ltd. Oe-FBW7 consisted of a pcDNA3.1 vector containing the full-length FBW7 cDNA sequence, with empty pcDNA3.1 as the negative control (Oe-NC).

Techniques: Control, Over Expression, Negative Control, Transfection

Journal: Experimental and Therapeutic Medicine

Article Title: F-box and WD repeat-containing protein 7 ameliorates angiotensin II-induced myocardial hypertrophic injury via the mTOR-mediated autophagy pathway

doi: 10.3892/etm.2022.11391

Figure Lengend Snippet: Schematic diagram presenting the hypothesis of the present study. FBW7, F-box and WD repeat-containing protein 7; Ang, angiotensin.

Article Snippet: The FBW7 overexpression plasmid (Oe-FBW7) and its negative control (Oe-NC) were obtained from Guangzhou RiboBio Co., Ltd. Oe-FBW7 consisted of a pcDNA3.1 vector containing the full-length FBW7 cDNA sequence, with empty pcDNA3.1 as the negative control (Oe-NC).

Techniques:

Journal: Experimental and Therapeutic Medicine

Article Title: Formononetin suppresses the malignant progression of papillary thyroid carcinoma depending on downregulation of CBX4

doi: 10.3892/etm.2024.12746

Figure Lengend Snippet: FMNT targets and downregulates CBX4 expression. (A) CBX4 expression levels in thyroid cancer tissues and normal tissues were obtained from the Encyclopedia of RNA Interactomes database. (B) CBX4 expression levels in TPC-1 cells and Nthy-ori3-1 cells were determined using reverse transcription-quantitative PCR and western blot analysis. *** P<0.001 vs. the Nthy-ori3-1 group. (C) Molecular docking of the compound-protein interactions between FMNT and CBX4. (D) TPC-1 cells were treated with 0, 10, 30 and 100 µM FMNT for 24 h. CBX4 expression levels in TPC-1 cells was determined by western blot analysis. *** P<0.001 vs. the Control group. FMNT, formononetin; CBX4, chromobox homolog 4.

Article Snippet: CBX4 overexpression plasmid (oe-CBX4) was established by inserting the CBX4 gene into a pcDNA3.1 vector (Shanghai GenePharma Co., Ltd.), whereas an empty vector served as the negative control (oe-NC).

Techniques: Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot, Control

Journal: Experimental and Therapeutic Medicine

Article Title: Formononetin suppresses the malignant progression of papillary thyroid carcinoma depending on downregulation of CBX4

doi: 10.3892/etm.2024.12746

Figure Lengend Snippet: FMNT treatment suppresses the proliferative and clone-forming abilities of papillary thyroid carcinoma cells by downregulating CBX4 expression. (A) TPC-1 cells were transfected with oe-CBX4 or oe-NC. CBX4 expression in TPC-1 cells was determined by reverse transcription-quantitative PCR and western blot analysis. *** P<0.001 vs. oe-NC. (B) FMNT-treated TPC-1 cells were transfected with oe-CBX4 or oe-NC. The clone-forming ability of TPC-1 cells was evaluated using a colony formation assay. *** P<0.001 vs. the Control group; # P<0.05 vs. FMNT + oe-NC. (C) FMNT-treated TPC-1 cells were transfected with oe-CBX4 or oe-NC. The proliferative ability of TPC-1 cells was evaluated using EdU staining (magnification, x200). FMNT, formononetin; CBX4, chromobox homolog 4; oe, overexpression; NC, negative control.

Article Snippet: CBX4 overexpression plasmid (oe-CBX4) was established by inserting the CBX4 gene into a pcDNA3.1 vector (Shanghai GenePharma Co., Ltd.), whereas an empty vector served as the negative control (oe-NC).

Techniques: Expressing, Transfection, Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot, Colony Assay, Control, Staining, Over Expression, Negative Control

Journal: Experimental and Therapeutic Medicine

Article Title: Formononetin suppresses the malignant progression of papillary thyroid carcinoma depending on downregulation of CBX4

doi: 10.3892/etm.2024.12746

Figure Lengend Snippet: FMNT treatment represses the migration, invasion, EMT and angiogenesis of papillary thyroid carcinoma cells by downregulating CBX4 expression. FMNT-treated TPC-1 cells were transfected with oe-CBX4 or oe-NC. (A) The migratory ability of TPC-1 cells was evaluated by wound healing assay (magnification, x100). (B) The invasive ability of TPC-1 cells was evaluated using a Transwell assay (magnification, x100). (C) Expression levels of MMP2 and MMP9 in TPC-1 cells were determined by western blotting. (D) Expression levels of E-cadherin, N-cadherin and Vimentin in TPC-1 cells were determined by western blot analysis. (E) HUVECs were incubated with the CM of TPC-1 cells at 37˚C for 24 h. In vitro angiogenesis of HUVECs was evaluated by tube formation assay (magnification, x200). (F and G) VEGF and VEGFR2 levels in TPC-1 cells were determined by reverse transcription-quantitative PCR. *** P<0.001 vs. the Control group; ## P<0.01 and ### P<0.001 vs. FMNT + oe-NC. FMNT, formononetin; CBX4, chromobox homolog 4; oe, overexpression; NC, negative control; HUVECs, human umbilical vein endothelial cells.

Article Snippet: CBX4 overexpression plasmid (oe-CBX4) was established by inserting the CBX4 gene into a pcDNA3.1 vector (Shanghai GenePharma Co., Ltd.), whereas an empty vector served as the negative control (oe-NC).

Techniques: Migration, Expressing, Transfection, Wound Healing Assay, Transwell Assay, Western Blot, Incubation, In Vitro, Tube Formation Assay, Reverse Transcription, Real-time Polymerase Chain Reaction, Control, Over Expression, Negative Control

Journal: Experimental and Therapeutic Medicine

Article Title: Formononetin suppresses the malignant progression of papillary thyroid carcinoma depending on downregulation of CBX4

doi: 10.3892/etm.2024.12746

Figure Lengend Snippet: FMNT treatment inhibits stemness characteristics of papillary thyroid carcinoma cells by downregulating CBX4 expression. FMNT-treated TPC-1 cells were transfected with oe-CBX4 or oe-NC. (A) Sphere formation of TPC-1 cells was evaluated by sphere formation assay (magnification, x200). (B) Expression levels of NANOG, OCT4 and CD133 in TPC-1 cells were determined by western blot analysis. *** P<0.001 vs. the Control group; ## P<0.01 and ### P<0.001 vs. FMNT + oe-NC. FMNT, formononetin; CBX4, chromobox homolog 4; oe, overexpression; NC, negative control.

Article Snippet: CBX4 overexpression plasmid (oe-CBX4) was established by inserting the CBX4 gene into a pcDNA3.1 vector (Shanghai GenePharma Co., Ltd.), whereas an empty vector served as the negative control (oe-NC).

Techniques: Expressing, Transfection, Tube Formation Assay, Western Blot, Control, Over Expression, Negative Control